ANALYSIS OF DETECTION OF HBV-NRAg IN SERUM AND ITS CORRELATION WITH HBV DNA

[目的]了解HBV携带者血清HBV-NRAg的检出情况,探讨HBV感染后HBV-NRAg与HBVM、HBVDNA载量的相关性。[方法]连续收集了从业人员的体检标本(n=4968),ELISA法检测血清HBV-NRAg和HBVM,同时以FQ-PCR检测核酸指标并辅以分区套式PCR和DNA测序作为证实检测,统计分析HBV-NRAg与HBVM、HBVDNA的相关性。[结果]HBV-NRAg的阳性率7.5%,略低于HBsAg的8.3%,略高于HBV DNA的6.8%。在HBV携带者中HBV-NRAg检测与HBV DNA的符合率87.4%,高于HBsAg或HBeAg,其S/CO值与HBVDNA载量线性相关(R2=0.927)。通过HBV-NRAg的检测筛选到6例HBV隐匿性感染,其中3例存在S区"a"基因突变。[结论]对于HBV感染后病毒复制状态和传染性的评价,HBV-NRAg可以替代HBV DNA作为良好的评判指标,而且可以用于变异毒株的筛选。[Objective]To investigate detection situation of HBV-NRAg in serum from HBV carriers,and discuss the correlation of the HBV-NRAg with HBVM and HBVDNA loads,respectively.[Methods]4968 serum samples were consecutively collected from the health examination of service workers,and HBV-NRAg and HBVM were detected by using ELISA.At the same time,the nucleic acid index of HBV DNA was examined by FQ-PCR,supplemented with detecting nested-PCR and DNA sequencing as certified tests.Statistical analyses were performed to analysis the correlation of HBV-NRAg with HBVM and HBVDNA,respectively.[Results]The total positive rate of HBV-NRAg was 7.5%,which was lower than HBsAg(8.3%) but higher than HBV DNA(6.8%) .A good conformability was observed between the test results of HBV-NRAg and HBV DNA.Meanwhile the S/CO values was closely correlated with HBV DNA loads(R2=0.927) .Furthermore,6 cases of concealed HBV infection were screened out by HBV-NRAg test,in which HBV genome had a point mutation in the‘a’epitope of S region.[Conclusion]As a replacement of HBV DNA,HBV-NRAg can be seen as a good evaluation index in terms of the evaluation on replicating phase HBV and its infectivity after being infected.Moreover,it also can be applied to effective screening of HBV mutation.厦门市重大疾病科研攻关项目(WKZ0501

南美白对虾肠道微生物群落的分子分析

采用分子生物学手段16S rDNA克隆文库方法对实验室养殖条件下的南美白对虾肠道细菌进行了多样性研究。用限制性片段长度多态性(RFLP)方法从文库中筛选出可能不同细菌来源的克隆子12个,测定其16S rDNA片段核甘酸序列,将所获得的序列与GenBank数据库进行BLAST比对,结果表明:南美白对虾肠道的16S rDNA克隆文库中126个克隆子分属2个不同的细菌类群:变形细菌(Proteobacteria)和厚壁细菌(Firmicutes),其中厚壁细菌为优势菌群占到75.4%,且与最相似序列同源性均低于94%;变形细菌占到24.6%,与最相似序列同源性均高于98%,分别为希瓦氏菌属(Shewanella),泛菌属(Pantoea),Aranicola属,假单胞菌属(Pseudomonas)和弧菌属(Vibrio)

Assay of biotoxicity of polycyclic aromatic hydrocarbons by Microtox test.

利用Microtox 技术检测5 种多环芳烃化合物生物毒性结果表明,二甲亚砜配制的测试液中萘、菲及荧蒽均对发光细菌具有一定生物毒性,且随浓度的增大而增强,相同浓度下毒性菲> 萘;测试液中当萘浓度小于其溶解度时即产生100%的抑光率,萘EC50为4.32mg/ L ,而菲及荧蒽浓度近其溶解度时所产生的最大抑光率分别为< 50 %和15 %左右;芘及蒽最大浓度时则对发光细菌无生物毒性显示。表明Microtox 技术可有效检测低环化合物萘的生物毒性,但对高环化合物( ≥3 环) 的检测因受其低水溶性的限制而灵敏度降低,利用二甲亚砜获取多环芳烃污染物提取液的生物毒性主要与低分子化合物萘及菲有关。国家自然科学基金项目(30070157) 、福建省教育厅基金项目(KB 0316) 和福建省泉州市科技计划重点项目(Z200234) 共同资

Microtox技术检测多环芳烃生物毒性的研究

利用Microtox技术检测 5种多环芳烃化合物生物毒性结果表明 ,二甲亚砜配制的测试液中萘、菲及荧蒽均对发光细菌具有一定生物毒性 ,且随浓度的增大而增强 ,相同浓度下毒性菲 >萘 ;测试液中当萘浓度小于其溶解度时即产生 10 0 %的抑光率 ,萘EC50 为 4 .32mg/L ,而菲及荧蒽浓度近其溶解度时所产生的最大抑光率分别为 <5 0 %和15 %左右 ;芘及蒽最大浓度时则对发光细菌无生物毒性显示。表明Microtox技术可有效检测低环化合物萘的生物毒性 ,但对高环化合物 (≥ 3环 )的检测因受其低水溶性的限制而灵敏度降低 ,利用二甲亚砜获取多环芳烃污染物提取液的生物毒性主要与低分子化合物萘及菲有

虾池沉积环境中若干功能菌及弧菌的时空变化

2003年10月～2004年5月在泉州东海一新建虾池与一池龄15年的老旧虾池,采用平板计数方法研究底泥环境中可培养异养菌(HB) 、淀粉降解菌(AB) 、有机磷降解菌OPB) 、无机磷溶解菌( INP) 、几丁质降解菌(CB) 、油脂降解菌(LB) 、纤维素降解菌(CLB) 、硫氧化细菌( SOB)等各种功能菌以及弧菌(VB)数量的变化情况,并对它们与可培养异养菌之间的相关关系进行了探讨. 结果表明,在整个养殖期中,新池泥样中可培养异养菌总数范围在1. 95 ×104 ～7. 7 ×105CFU /g之间, 旧池泥样中可培养异养菌总数范围在2 ×104 ～1. 88 ×105 CFU /g之间,两池的数量变化波动均较大,其它功能菌与异养菌相似,在整个养殖周期也是呈现较大的波动幅度, 但统计分析表明淀粉降解菌、有机磷降解菌、油脂降解菌、弧菌等与可培养异养细菌之间呈现着明显的正相关,而几丁质降解菌、无机磷溶解菌、纤维素降解菌以及硫氧化细菌与异养菌之间则无明显的相关关系.国家863项目(2002AA603013) ,国家自然科学基金项目 (30370276) ,福建省科技重点项目(2004 I023)资

海水养殖沉积环境微生物总DNA的提取方法研究

传统的微生物分离与培养技术无法揭示微生物群落的动态变化.为了阐释虾池沉积环境微生态格局的原始组成情况,本研究先以TENP缓冲液去除沉积物中的腐殖酸,继之以溶菌酶-SDS温和裂解,其总DNA的提取效率达90%以上,所获DNA产量达2~20μg/g沉积物(湿),片段大小均在23 kb左右,不经纯化即可直接进行PCR扩增和限制性酶切.以该DNA为模板进行PCR-DGGE分析,揭示了虾池沉积物丰富的微生物多样性.该方法是一种适用于虾池沉积物总DNA提取的简便、可靠方法

阳离子聚合物介导下白细胞介素1受体拮抗剂基因兔角膜原位转染及其表达

目的探讨阳离子聚合物即线性聚乙烯亚胺(PEI)介导下兔角膜基质注射PEGFP-IL- 1ra质粒进行基因角膜原位转染的有效性和安全性。方法以人cDNA文库为模板进行聚合酶链反应(PCR),获得人IL-1ra cDNA片段,构建重组质粒PEGFP-hIL-1ra。以阳离子聚合物为介导转染角膜内皮细胞,通过绿色荧光蛋白(GFP)示踪、蛋白免疫印迹技术检测转染后IL-1ra基因和蛋白的表达。实验组30只Wistar大鼠角膜基质注射PEGFP-hIL-1ra质粒和PEI-in-vivo的混合溶液20μl(含10μg质粒),对照组15只Wistar大鼠角膜基质注射PEI-in-vivo溶液20μl。注射后1、3、6、14、21 d,收集角膜通过HE染色、透射电镜、锥虫蓝-茜素红染色、免疫组织化学观察1L-1ra基因角膜原位转染后的细胞结构和功能的变化。荧光显微镜下追踪IL-1ra-GFP融合蛋白在角膜的表达部位和表达强度。结果以cDNA文库为模板扩增出hIL-1ra cDNA片段,构建重组质粒PEGFP-hIL-1ra。经PstI和BamHI酶切及DNA测序证实了插入片段方向和大小正确。PEI-in-vitro介导下PEGFP-hIL-1ra转染角膜内皮细胞12 h后,可见10％-15％细胞中有GFP荧光表达。Western-blotting检测可见相对分子质量为44 000的hIL-1ra-GFP蛋白表达。PEGFP-hIL-1ra质粒和PEI-in-vivo角膜基质注射后1 d,角膜上皮基底细胞可见荧光条带,6 d时全角膜荧光强度达到高峰,14 d开始减弱,21d角膜上皮层尚存微弱荧光。对照组观察期内始终未见绿色荧光。实验组角膜HE染色未见病理性改变,角膜上皮基底细胞层p63抗体阳性;锥虫蓝-茜素红联合染色未见角膜内皮细胞损伤;透射电镜显示角膜各层细胞的细胞质内可见IL-1ra-GFP颗粒,未见细胞器的损害。结论阳离子聚合物介导下角膜基质注射PEGFP-hIL-1ra质粒可快速、有效地将IL-1ra基因转入角膜并表达,为临床上使用抗炎细胞因子IL-1ra对角膜免疫炎性反应相关疾病进行基因治疗提供了新的技术平台

C–X(X = Cl, Br, I) bond dissociation energy as a descriptor for the redispersion of sintered Au/AC catalysts

负载型Au基催化剂在工业过程中具有非常广泛的潜在应用,如催化加氢/脱氢过程、精细化学品合成、能源催化转化及环境保护等过程,表现出很高的催化活性和选择性.Au基催化剂活性物种或活性中心基本由纳米粒子或化合物构成,但在应用过程中因Ostwald熟化效应或粒子迁移作用,尤其是高温高压等苛刻反应条件下,均随应用时间延长从小尺寸粒子逐渐长为大粒子,造成活性降低或完全失活,这也是负载型催化剂失活的最主要原因之一.其中因成本、稀缺等特性,负载型Au催化剂的烧结问题是影响和制约其应用的主要因素.除可通过载体改性、助剂和官能团配位稳定等方法来延缓其失活过程外,对已烧结催化剂的高效、快捷和绿色的再分散/再生过程也具有基础和应用研究的重要意义.活性炭载Au催化剂(Au/AC)广泛应用于乙炔氢氯化反应中,以期替代高毒性的汞基催化剂,但在反应过程中因高活性的Au~(3+)物种易被还原而形成Au~0物种进而烧结导致失活;如新鲜Au/AC催化剂表面的Au粒子尺寸为1-2 nm,经乙炔氢氯化反应后变为33 nm左右;随之在453 K、0.1 MPa、乙炔体积空速(GHSV)为600 h~(-1)、氯化氢与乙炔摩尔比为1.1的反应条件下,乙炔转化率从81.8%降至11.2%.如何有效对大粒子Au再分散/再生可为其应用提供有力支撑.有研究表明,气相CHI_3在甲醇羰基化反应过程中明显改变Au/AC表面的Au粒子尺寸;或采用浓盐酸或王水也可将烧结的Au/AC催化剂进行再分散/再生.但已有的Au基催化剂再分散/再生过程均伴随着强酸、强氧化或高毒性在分散剂的应用,对环境的影响及后续处理有明显的局限性,且再分散机理尚不明确.在前期工作基础上,本文采用系列卤代烃(碘代烃、溴代烃和氯代烃)对烧结的Au/AC进行再分散/再生研究.结果表明,在室温常压条件下CHI_3可以快捷高效地对烧结Au/AC催化剂进行再分散/再生,具有最优的再分散性能;通过对系列碘代烃C-I键的解离能分析,发现C-I解离能越低越有利于大粒子Au的再分散.同时,溴代烃和氯代烃对烧结的Au/AC催化剂也具有再分散能力,但比碘代烃的再分散效率低.C-X键的解离能与再分散效率有高相关性,即C-X键的解离能越低越有利于Au的再分散.总体上,三类卤代烃再分散效率高低顺序为C-I>C-Br>C-Cl.进而,通过不同分散过程中Au粒子分散状态推测了卤代烃对Au粒子的再分散机理,即卤代烃先在Au粒子表面化学吸附,然后C-X键解离,形成Au-X物种,小粒子Au在AC表面聚集并稳定,最后形成高分散Au粒子(粒径<1 nm)催化剂.以乙炔氢氯化反应考察了再生Au/AC催化剂性能,结果表明,该催化剂上乙炔转化率可达79.4%,基本恢复至初始水平,且该方法可对失活催化剂进行多次高效再生.Disintegration or redispersion of supported sintered gold nanoparticles(Au NPs) in the presence of alkyl halide can give catalyst regeneration or redispersion of sintered Au catalysts. The selectivity of alkyl halides, temperature and size distributions were investigated to elucidate the redispersion of Au NPs during halide-induced decomposition. This study proved that the alkyl halide induced the redispersion of sintered Au NPs which depended on the R–X(X = I, Br, Cl) bond dissociation energy(BDE) and thus provided a simple descriptor for the regeneration of inactive supported Au cata-lysts. A correlation between the BDE of R–X and dispersion efficiency was established. The tendency for disintegration and redispersion followed the R–X BDE of the alkyl halide. Compared to alkyl chlorides and bromides, iodides were more efficient for redispersing sintered Au NPs. As a descriptor, the BDE of R–I played a crucial role in particle redispersion. These findings provided insights into the mechanism of organic halide-induced Au NP disintegration and the effect of the halide type on the redispersion of sintered catalysts.国家自然科学基金(21403178,21473145,21503173,91545115);; 教育部创新团队发展计划(IRT_14R31);; 福建省青年教师教学科研基金(JA15003

Preliminary study on PAH degradation by bacteria from contami-nated sediments in Xiamen Western Sea, Fujian, China

In order to estimate the biodegradation of three polycyclic aromatic hydrocarbons (PAHs) compounds, bacterial strains were isolated from marine sediments in three heavily contaminated sites (Yuandang Lake, Dongdu Port and Aquacultural zones in Maluan Bay) in Xiamen Western Sea. The results show three bacterial strains, which used pyrene as the sole carbon source, were identified as strains of Aureobacterium sp., Arthrobacter sp., Rhodococcus sp. The PAH-degrading bacteria isolated had a strong ability to degrade phenan-hrene, fluoranthene and pyrene at different degradation rates. The highest degradation rate was observed when three PAH compounds were mixed with an individual strain in the medium. The three PAHs were degraded after one week with a degradation rate of 89.94 % for phenanthrene and 93.4 % for both of fluoranthene and pyrene. In addition, after 25 days of incubation, the degradation rate was 99.98 % for phenanthrene and 99.97 % for both of fluoranthene and pyrene. Optical density was measured to ..

Phytoremediation of heavy metals and its endophytic bacteria effects

土壤和水体的重金属污染已严重危害人类生存环境与健康。由于受重金属污染的环境分布广泛,迫切需要开发经济的清除环境重金属的技术。植物修复是通过绿色植物降解或移除环境污染物,有望成为重金属污染环境的原位修复技术。植物内生菌是指定殖于健康植物的各种组织和器官内部的细菌,被感染的宿主植物不表现出外在病症,耐重金属的内生菌在多种超富集植物中存在。在植物修复过程中,野生型内生菌或基因工程内生菌的抗性系统能降低重金属植物毒性,促进其迁移金属。耐重金属内生菌还可以通过固氮、溶解矿物元素及产生类植物激素、铁载体和ACC脱氨酶等产物促进植物的生长。主要综述目前植物-内生菌相互作用及其潜在的促进植物修复重金属污染的研究进展。Pollution of soils and water with heavy metals is becoming one of the most severe environ-mental and human health hazards.Innovative ways are pressed for cleaning widespread heavy metals contamination.Phytoremediation is a remediation technology that requires the use of green plants to remove pollutants from the environment.Endophytic bacteria colonize within plant hosts without caus-ing symptoms of infection or negative effects.The metal resistant endophytes are present in various hyperccumulatorplants.During phytoremediation of metals,the wild-type or engineered endophytes can lower metal phytotoxicity and enhance heavy metal translocation to plant through its metal-resistance system.Moreover,the metal resirtant endophytes can indirectly benefit plant growth by various mecha-nisms such as nitrogen fixation,solubilization of minerals production of phytohormines,siderophors,1-aminocyclopropane-1-carboxylate(ACC) deaminase.This review describes the potential for exploit-ing plant-endophyte pertnerships to improve phytoremediation of heavy metals.国家自然科学基金项目(No.31070296);福建省高校服务海西建设重点项目(No.A102